产脂肪酶菌株C7828-5的筛选、鉴定以及产酶条件的优化

以花生油为唯一碳源,从海口市各地被油脂污染土样中分离筛选出1株中温碱性脂肪酶菌株C7828-5。形态学、生理生化特征和分子生物学鉴定结果表明,该菌株为铜绿假单胞菌(Pseudomonas aeruginosa)。该菌所产脂肪酶的最适温度为37℃,最适pH为8.0。优化了菌株的产酶条件,最适产酶培养基(g/L)为:蔗糖5、牛肉膏20、(NH_4)_2SO_41、MgSO_4·7H_2O 0.5、CaCl_20.5,聚乙烯醇花生油乳化液120 mL,发酵72 h,获得高达8.08 U/mL的脂肪酶表达量。     

铜绿假单胞菌产蛋白酶的发酵条件优化

【目的】鉴定一株来源于酱油曲能够分泌蛋白酶的铜绿假单胞菌CAU342A,优化其产蛋白酶的发酵条件。【方法】采用形态学观察、16S r RNA基因序列比对和生理生化方法鉴定菌株CAU342A;通过碳源、氮源、初始pH、温度、表面活性剂及发酵时间的单因素优化和正交试验获得最适发酵条件。【结果】菌株CAU342A被鉴定为铜绿假单胞菌(Pseudomonas aeruginosa),其最适发酵产酶条件为(质量体积比):3%酒糟,1.5%酵母浸提物,0.05%吐温-80,0.5%NaCl,0.7%K_2HPO_4,0.3%KH_2PO_4,0.04%MnSO_4,培养基初始pH 7.5,30°C培养72 h。在最适发酵条件下,该菌株最大产酶水平达到2 653.5 U/m L。蛋白酶酶谱分析表明该菌株能够产生至少4种具有蛋白酶活性的同工酶,其中两个主要酶谱带对应分子量分别为32 k D和50 k D。【结论】铜绿假单胞菌CAU342A高产蛋白酶,具有很大的工业应用潜力。     

产脂肪酶嗜碱细菌的筛选及酶学性质研究

目的:筛选产脂肪酶嗜碱细菌,并研究其酶学性质.方法:以豆油为唯一碳源的固体平板筛选产酶菌株,16S rDNA同源性分析确定微生物菌属,单因素实验优化产酶条件、研究酶学性质.结果:筛选出1株产脂肪酶的嗜碱菌株,鉴定为假单胞菌(Pseudomonas),命名为Pseudomonas sp.C-36.菌株产酶的最佳培养条件为:甘油2%(V/V).蛋白胨0.7%(W/V),酵母提取物0.5%(W/V),K2HPO4 0.2%(W/V),MsS04·7H2O 0.05%(W/V),NaCl 0.3%(W/V),Triton X-100 0.01%(W/V),pH 9.5,转速180r/min,37℃下培养24h,产酶量为2.782 IU/ml.该酶的最适温度和pH分别为40℃和9.0,50℃时酶活半衰期为2h.Ca2+等金属离子对该酶酶活具有促进作用,而Zn2+对酶活的抑制作用明显.有机溶剂的耐受性实验表明,该酶在疏水性有机溶剂中稳定性良好.结论:筛选得到1株嗜碱的脂肪酶产生菌C-36,并鉴定为Pseudomonas.     

假单胞菌产脂肪酶条件的初步探索

对假单胞菌（Pseudomonassp）2106菌株产脂肪酶条件的初步探索表明,该菌株脂肪酶为组成型,不受油脂类底物的诱导。碳源的种类（单糖、双糖、多糖）和浓度对产酶影响不大,氮源以豆饼粉和玉米浆混合添加最好。最适发酵温度为32℃,摇瓶转速140r／min。实验中还通过正交试验优化了产酶培养基组成。     

壳聚糖酶生产菌的筛选、鉴定及其产酶培养条件的研究

从土样中分离到60株分泌胞外壳聚糖酶的菌株,经过筛选,其中有1株细菌产酶能力较高.生理生化试验鉴定该细菌为假单胞菌(Pseudomonas sp.).对假单胞菌产酶的培养条件研究结果表明:最适培养基组分为(g/L):壳聚糖5,氨基葡萄糖2,硝酸氨2,MgSO4·7H2O 0.5,KH2PO4 0.4,KCL 0.5,FeSO4·7H2O 0.01,起始pH 6.5;适宜培养条件是:接种量2.0×107个/50ml培养液,28℃,120 r/min振荡培养3d.     

荧光假单孢菌Pseudomonas fluorescence 5963 产脂肪酶条件的优化

对荧光假单孢菌Pseudomonas fluorescence 5963产脂肪酶条件进行了筛选.该菌株的最适产酶条件如下(w/v)1%淀粉作为碳源;2%酵母抽提物作为氮源;0.03%Mg2SO4·7H2O;0.2%诱导物,水1 L;pH 7.0;培养温度为28℃.     

利用麻风树油驯化筛选高活性脂肪酶产生菌及其催化酯化反应

以1株分解麻风树油的脂肪酶产生菌Pseudomonas sp. LP-1为出发菌株, 通过麻疯树油定向驯化筛选获得1株酶活较高且产酶稳定的菌株P. sp. X-2-45, 其水解酶活为29.79 U/mL, 比原始菌株提高了288%。对P. sp. X-2-45生长与产酶特征、对植物油脂水解能力及在有机相中催化脂肪酸和有机醇间的酯化反应研究发现, 该菌株生长速率和产酶速率明显加快, 培养30 h时生物量和酶活达到最大, 稳定期延长, 培养过程中脂肪酶在培养基中的稳定性提高。以麻疯树油诱导合成的P. sp. X-2-45脂肪酶对麻疯树油的水解能力比原始菌株提高了378%, 说明采用麻风树油定向驯化可提高脂肪酶对相应底物的水解能力。X-2-45脂肪酶可以催化月桂酸与正丁醇、正辛醇、月桂醇和丙三醇之间, 棕榈酸、硬脂酸与甲醇、正辛醇、月桂醇和丙三醇之间, 油酸与甲醇、正丁醇、正辛醇、月桂醇和丙三醇之间发生酯化反应。     

黑色葡萄状穗霉S607耐碱性纤维素酶发酵条件的研究

研究了耐碱性纤维素酶生产菌株黑色葡萄状穗霉S6 0 7(StachybotrysatraS6 0 7)的产酶条件。结果表明 ,S6 0 7的最适生长和产酶 pH为 7.0 ,最适产酶温度为 2 8℃ ,摇床最适转速为 180r/min。在以 2 .0 %纤维素粉和 2 .5 %的麸皮作为碳源时产酶最高 ,添加含 (NH4 ) 2 SO4 、尿素和蛋白胨的复合氮源对生长和产酶有明显的促进作用。在最适培养条件下 ,发酵周期为 4～ 5d ,发酵液中CMC酶活为 2 .12IU /mL ,FP酶活为 1.0 3IU /mL ,β 葡萄糖苷酶活为 0 .86IU /mL。     

聚乙烯醇降解酶产生菌的分离和发酵条件

从工厂废水中分离到一株高效降解聚乙烯醇(PVA)的细菌D8株,四天能将培养基中0.5％的PVA(500,1700)完全降解,经鉴定为假单胞菌属类产碱假单胞菌(Pseudoraonospseudoalcaligenes)。对菌株的发酵条件进行了研究,结果表明,最适培养基成份为(％)：PVA1．5,(NH4),SO4 0．1,K2HPO4 0．24,KH2PO4 0．04, MgSO4·7H2O 0.035,NaCl 0.01,FeSO4 0.001,酵母膏0.15;起始pH 7.5;通气量在250ml三角瓶装30ml培养基为最逢,于30℃,160r／rain的旋转摇床振荡培养72h产酶活力最高。     

交替假单胞菌LP621菌株产右旋糖苷酶的培养条件优化

从江苏连云港海域分离和筛选到1株产右旋糖苷酶的海洋细菌交替假单胞菌Pseudoalteromonas tetraodonis LP621,通过单因素试验和正交试验对该菌株产右旋糖苷酶培养条件进行优化。单因素试验结果表明,最佳培养时间为24 h,最适产酶温度为25℃;产酶pH范围为5.0~11.0,最适产酶pH为6.0;产酶NaCl浓度范围为1%~10%,NaCl浓度为4%时产酶较高;装液量在25%。麦芽糖、胰蛋白胨和酵母膏促进产酶。利用响应面方法对LP621产右旋糖苷酶的发酵条件进行优化。选择培养基pH、时间、麦芽糖浓度和装液量4因素进行优化,结果为pH 7.07,发酵时间21.94 h,麦芽糖浓度0.42%,装液量为21.88%,酶活为270.1U/mL。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.